Binding reagents that contain small epitope binding molecules

Abstract

The invention provides binding reagents that contain a plurality of linked small epitope binding molecules that each recognizes a small epitope, such as small epitope antibodies. The combination of small epitope binding molecules in a binding reagent specifically recognizes and binds to a molecule of interest. The binding reagents may be used for such purposes as detection, quantification, identification, and purification of molecules of interest.

1 . A binding reagent comprising a plurality of small epitope binding molecules, wherein said small epitope binding molecules are linked, and wherein each of said small epitope binding molecules recognizes a different small epitope. 
   
   
       2 . A binding reagent according to  claim 1 , wherein said plurality of small epitope binding molecules consists of 2, 3, or 4 small epitope binding molecules. 
   
   
       3 . A binding reagent according to  claim 1 , wherein said small epitope binding molecules are small epitope antibodies or epitope binding fragments thereof. 
   
   
       4 . A binding reagent according to  claim 3 , wherein each small epitope antibody or epitope binding fragment thereof recognizes an epitope consisting of 3, 4, or 5 contiguous amino acids. 
   
   
       5 . A binding reagent according to  claim 3 , wherein said plurality of small epitope binding molecules consists of 2, 3, or 4 small epitope antibodies or epitope binding fragments thereof. 
   
   
       6 . A binding reagent according to  claim 1 , wherein said small epitope binding molecules are selected from the group consisting of aptamers, molecular imprints, lectins, and capture compounds. 
   
   
       7 . A binding reagent according to  claim 1 , wherein the binding reagent binds specifically to a protein. 
   
   
       8 . A binding reagent according to  claim 1 , wherein the small epitope binding molecules are linked covalently. 
   
   
       9 . A binding reagent according to  claim 8 , wherein the covalent linkage comprises a chemical cross-linking reagent. 
   
   
       10 . A binding reagent according to  claim 1 , wherein the small epitope binding molecules are linked by interaction between members of a binding pair. 
   
   
       11 . A binding reagent according to  claim 3 , wherein the reagent is recombinantly expressed as a polypeptide with a peptide linker between the antibodies or epitope binding fragments. 
   
   
       12 . A binding reagent according to  claim 1 , comprising a detectable label. 
   
   
       13 . A library of binding reagents, comprising a plurality of binding reagents according to  claim 1 , wherein each of said binding reagents recognizes a different combination of small epitopes. 
   
   
       14 . A method of characterizing a molecule of interest, said method comprising contacting a sample suspected of comprising the molecule of interest with a binding reagent according to  claim 1 , wherein said binding reagent is capable of specifically binding to the molecule of interest, and detecting binding of the molecule of interest to the binding reagent. 
   
   
       15 . A method according to  claim 14 , wherein characterizing comprises determining the amount of the molecule of interest in the sample. 
   
   
       16 . A method according to  claim 14 , wherein characterizing comprises determining presence or absence of the molecule of interest in the sample. 
   
   
       17 . A method according to  claim 14 , wherein the molecule of interest is a biomarker. 
   
   
       18 . An array comprising a plurality of binding reagents according to  claim 1  immobilized on a support, wherein each of said binding reagents recognizes a different combination of small epitopes. 
   
   
       19 . A method of making a binding reagent according to  claim 1 , comprising linking said small epitope binding molecules with a chemical cross-linking reagent. 
   
   
       20 . A method of making a binding reagent according to  claim 1 , comprising linking said small epitope binding molecules via interaction between members of a binding pair. 
   
   
       21 . A method of making a binding reagent according to  claim 3 , comprising recombinant expression of a polypeptide comprising said small epitope antibodies or epitope binding fragments thereof. 
   
   
       22 . A kit comprising a binding reagent according to  claim 1 . 
   
   
       23 . A composition comprising a binding reagent according to  claim 1 . 
   
   
       24 . A pharmaceutical composition comprising a binding reagent according to  claim 1  and a pharmaceutically acceptable carrier. 
   
   
       25 . An affinity matrix comprising a binding reagent according to  claim 1  attached to a substrate. 
   
   
       26 . A method for purifying a molecule of interest, comprising contacting an affinity matrix according to  claim 25  with a sample comprising the molecule of interest, wherein said binding reagent binds to said molecule of interest. 
   
   
       27 . A method according to  claim 26 , wherein said molecule of interest is selected from the group consisting of a polypeptide, a polynucleotide, a polysaccharide, and a glycoprotein.